土壤微生物总 DNA 提取及其 PCR 优化 |
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引用本文: | 李超,梁俊峰.土壤微生物总 DNA 提取及其 PCR 优化[J].桉树科技,2015(2):17-21. |
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作者姓名: | 李超 梁俊峰 |
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作者单位: | 1. 中国林业科学研究院热带林业研究所,广东 广州 510520; 国家林业局桉树研究开发中心,广东 湛江 524022;2. 中国林业科学研究院热带林业研究所,广东 广州,510520 |
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基金项目: | 中国林业科学研究院基本科研业务费专项,广东省林业科技创新重点项目 |
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摘 要: | 采用3种不同土壤微生物总 DNA 提取方法对广东省乐昌杨东山十二度水自然保护区的样地土壤进行比较研究,进一步通过 PCR 扩增,研究牛血清白蛋白(BSA)对整个 PCR 扩增过程的影响。结果表明:试剂盒提取法相对于其他两种提取方法更加方便有效,当土壤样品量比较大时,直接提取法是最经济有效的方法;而在土壤微生物的 PCR 扩增实验中,加入2μl 的 BSA,可以有效地抑制腐殖酸等对后续 PCR 扩增的影响。
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关 键 词: | 土壤微生物 DNA提取 牛血清白蛋白 PCR扩增 |
Comparison of Extraction Methods For Soil Microbe DNA and Effectiveness of BSA for PCR Amplification |
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Abstract: | The extracting efficiency of three methods for soil DNA extraction and isolating of microbial soil DNA from sites in Yangdongshan Shierdushui Nature Reserve was examined. Also, the effect of bovine serum albumin (BSA) on PCR amplification of the DNA obtained was studied. The results showed that the DNA Extraction Kit method was the most simple and effective method. However, for large numbers of soil samples, the most economical method proved to be ‘the direct method’. It was found that 2 μl BSA could inhibit the deleterious effects of humic acid on PCR amplification of soil microbe DNA. |
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Keywords: | soil microbes DNA extraction BSA PCR amplification |
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